This project will use a team-based approach to pursue three directly interrelated areas of investigation important for learning to make new pancreatic beta cells from other cell types.
In Aim 1 we seek to gain a deeper understanding of the molecular events that dictate formation of multiple endocrine cell types during pancreas development. This will be achieved by isolating a series of highly purified progenitor cell populations, performing digital gene expression analysis, and developing bioinformatics strategies for characterizing the differences among discrete cellular populations that will be temporally or genetically informative.
In Aim 2 we will seek to utilize the knowledge gained in Aim 1 to determine the veracity of protocols being used to direct the differentiation of hESCs towards pancreatic cell fates and to rationally improve these protocols by monitoring the expression of gene clusters that are specifically activated or repressed during mouse development.
In Aim 3 we will develop new mouse lines in which the expression of three transcription factors that have been reported to be capable of transdifferentiating pancreatic acinar to beta cells can be easily modulated by the administration of doxycycline. These mice will be used to explore how transdifferentiation actually occurs and to determine the extent to which the newly generated beta cells are functional and exhibit a gene expression profile similar to authentic beta cells. This project is based on the premise/hypothesis that multiple gene regulatory networks, which are normally set up during mouse development, must be established during directed or trans-differentiation of other cell types to achieve the beta cell-like functionalities necessary for clinical use. Greater knowledge of pancreas-specific gene regulatory networks, how they are established and differ among related cell populations, and determining whether they are present or absent in experimentally-derived cellular populations, will serve as a platform both for new discovery and protocol improvements. The six investigators in this project have a track record of productive collaborative interactions and bring specific knowledge and abilities necessary to accomplish these goals. Thus, it is anticipated that this project will generate important resources that will not only advance two of the overarching goals of the Beta Cell Biology Consortium but also have broad scientific impact and utility.

Public Health Relevance

Type 1 and Type 2 diabetes are diseases that cause significant morbidity and mortality and thus have an adverse economic impact. Both diseases are characterized by the destruction or dysfunction of insulin-secreting pancreatic beta cells. This application seeks to gain key information for developing new, cell-based replacement therapies that hold promise for achieving better glucose control than is currently possible.

Agency
National Institute of Health (NIH)
Institute
National Institute of Diabetes and Digestive and Kidney Diseases (NIDDK)
Type
Research Project--Cooperative Agreements (U01)
Project #
5U01DK089523-04
Application #
8522192
Study Section
Special Emphasis Panel (ZDK1-GRB-G (M3))
Program Officer
Sato, Sheryl M
Project Start
2010-09-20
Project End
2015-06-30
Budget Start
2013-07-01
Budget End
2014-06-30
Support Year
4
Fiscal Year
2013
Total Cost
$1,243,572
Indirect Cost
$437,720
Name
Vanderbilt University Medical Center
Department
Physiology
Type
Schools of Medicine
DUNS #
004413456
City
Nashville
State
TN
Country
United States
Zip Code
37212
Worchel, Hannah N; Magnuson, Mark A (2014) Cytokine-driven beta-cell production in vivo. Nat Biotechnol 32:63-4
Osipovich, Anna B; Long, Qiaoming; Manduchi, Elisabetta et al. (2014) Insm1 promotes endocrine cell differentiation by modulating the expression of a network of genes that includes Neurog3 and Ripply3. Development 141:2939-49
Clark, Jessica K; O'keefe, Ashley; Mastracci, Teresa L et al. (2014) Mammalian Nkx2.2+ perineurial glia are essential for motor nerve development. Dev Dyn 243:1116-29
Magnuson, Mark A; Osipovich, Anna B (2013) Pancreas-specific Cre driver lines and considerations for their prudent use. Cell Metab 18:9-20
Lugani, Francesca; Arora, Ripla; Papeta, Natalia et al. (2013) A retrotransposon insertion in the 5' regulatory domain of Ptf1a results in ectopic gene expression and multiple congenital defects in Danforth's short tail mouse. PLoS Genet 9:e1003206
Arnes, Luis; Leclerc, Kevin; Friel, Jessica M et al. (2012) Generation of Nkx2.2:lacZ mice using recombination-mediated cassette exchange technology. Genesis 50:612-24
Potter, Leah A; Choi, Eunyoung; Hipkens, Susan B et al. (2012) A recombinase-mediated cassette exchange-derived cyan fluorescent protein reporter allele for Pdx1. Genesis 50:384-92
Mastracci, Teresa L; Wilcox, Crystal L; Arnes, Luis et al. (2011) Nkx2.2 and Arx genetically interact to regulate pancreatic endocrine cell development and endocrine hormone expression. Dev Biol 359:1-11